CN1465977A - Occult blood gold standard test strip and preparation method thereof - Google Patents
Occult blood gold standard test strip and preparation method thereof Download PDFInfo
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- CN1465977A CN1465977A CNA021146497A CN02114649A CN1465977A CN 1465977 A CN1465977 A CN 1465977A CN A021146497 A CNA021146497 A CN A021146497A CN 02114649 A CN02114649 A CN 02114649A CN 1465977 A CN1465977 A CN 1465977A
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Abstract
The invention refers to an occult blood golden mark test strip and the making method, including the scaleboard and protection layer; the hand-held end and test end water-absorbing layers made of the filter paper, which are pasted two ends of the scaleboard, respectively; the detecting layer made set on the scaleboard between the two water-absorbing layers; the anti-human hemoglobin golden mark antibody layer set between the test end water-absorbing layer and detecting layer; the detecting and control lines set on the detecting layer; the detecting layer enveloped by the detecting and control lines which are formed by anti-human hemoglobin monomer, sheep and rabbit anti-human hemoglobin multiantibody, sheep anti-rat and rabbit IgG antibody and rabbit sheep IgG antibody.
Description
Technical field
The present invention relates to a kind of occult blood gold standard test strip and preparation method thereof.
Background technology
Stool occult blood test is the main diagnosis index of hemorrhage of digestive tract and tumour.At present clinically, the sieve test method that stool occult blood test is commonly used is the heme that utilizes in the Main Ingredients and Appearance haemoglobin (Hb) of blood, has similar peroxidase effect and the simple and easy chemo-assay that designs.The oxidized color developing agent of these class methods is more, as biphenylamine, neighbour-toluidine, o-tolidine, guaiaci lignum, phenolphthalin, aminopyrin, colourless peacock green, tetramethyl benzidine, diphenylamine, dimethylbenzidine etc.; Thereby method is numerous.Though said method is simple and easy to do, specificity is lower, disturbing factor is more.Animal blood, meat, liver and be rich in chlorophyll food, chalybeate, vitamin C, Chinese medicine etc. and all can cause false positive.
Of the present invention open
Fundamental purpose of the present invention is for a kind of occult blood gold standard test strip is provided, this test-strips is based on the target material human hemoglobin monoclonal antibody in the occult blood or how anti-, a kind of high specificity that adopts immunochromatography technique and collaurum colour developing principle and design, highly sensitive, and speed is fast, easy and simple to handle, need not the single stage method occult blood gold standard test strip of specialized facilities.
Another object of the present invention is for a kind of preparation method of occult blood gold standard test strip is provided, and this method is simple, stability, good reproducibility.
Fundamental purpose of the present invention can realize by following measure:
A kind of occult blood gold standard test strip comprises liner plate and protective seam; Be pasted with handheld terminal water accepting layer and the test lead water accepting layer of making by filter paper respectively at the two ends of liner plate; On the liner plate between handheld terminal water accepting layer and the test lead water accepting layer, be provided with the detection layers that cellulose membrane is made; Between test lead water accepting layer and detection layers, be provided with antihuman hemoglobin gold labeling antibody layer; On detection layers, be provided with detection line and control line.
Another object of the present invention can realize by following measure:
A kind of preparation method of occult blood gold standard test strip comprises golden labeling antibody layer and detection layers and goes up the preparation of wrapping tested survey line and control line, and the assembling of test-strips.
The preparation of described antihuman hemoglobin gold labeling antibody layer comprises: the preparation of i collaurum, in concentration is that the 1.2-1.8%, the concentration that add its volume in the gold chloride of 0.01-0.03% is 1% trisodium citrate, boiled 10-20 minute, and be reduced into the collaurum atom liquid of 15-50 nanometer; Ii antihuman hemoglobin antibody colloidal gold mark adds 2-10mg antihuman hemoglobin antibody and comprises monoclonal antibody, sheep rabbit antihuman hemoglobin resist more in the 100ml colloidal gold solution; In above-mentioned colloidal gold solution, press 0.1-0.6g/100ml again and add animal serum albumin; In above-mentioned colloidal gold solution, press the saline solution of 6-12ml/100ml adding 10% then again; The last centrifugal precipitation of going, get supernatant centrifugal again sediment, sediment is dissolved in by 4-10ml/100ml 0.02M, PH7.4Tris-HCl contain the 0.1-0.6% animal serum albumin, 0.02% Sodium azide gets antihuman hemoglobin antibody colloidal gold solution; Iii is impregnated with glass fibre or nonwoven fabrics with antihuman hemoglobin antibody colloidal gold solution, and the immersion amount is with till immersing liquid and beginning outwards to flow out, then at 37 ℃ of dry down golden labeling antibody layers.
The preparation of described antihuman hemoglobin antibody is that the spleen cell and the rat bone marrow tumour cell that adopt routine techniques will stem from the mouse of using purifying antihuman hemoglobin immunity inoculation merge, and selects to produce the hybridoma of required antihuman hemoglobin monoclonal antibody; Again hybridoma cell clone is got monoclonal antibody.
How anti-the preparation of described detection layers be to add fixing agent in and sheep anti mouse, rabbit igg antibody, the anti-sheep IgG of the rabbit antibody at antihuman hemoglobin monoclonal antibody, sheep, rabbit antihuman hemoglobin, and how anti-and sheep anti mouse, rabbit igg antibody, the anti-sheep IgG of rabbit antibody be sprayed on and form detection line and control line on the cellulose membrane with antihuman hemoglobin monoclonal antibody, sheep, rabbit antihuman hemoglobin with Membrane jetter then; Contain 10% calf serum sealing rinsing in 30 minutes, the dry detection layers that gets with 0.01MPH7.0PBS liquid again.
Described fixing agent is selected from a kind of in formaldehyde, the acetone.
The present invention has following advantage compared to existing technology:
Adopt single stage method when 1, gold test strip bar of the present invention detects occult blood, have the specificity and the high sensitivity of height, it detects fast, and the result judges and finished in 3-15 minute, need not dedicated experiments facility, test condition and special operating personnel.
2, the preparation method of test strips of the present invention have simple for production, stability, good reproducibility.
The drawing explanation
Fig. 1 is a structural representation of the present invention
1-liner plate 2-protective seam 3-handheld terminal water accepting layer 4-detection layers
5-control line 6-detection line 7-antihuman hemoglobin gold labeling antibody layer
8-test lead water accepting layer
Embodiments of the present invention
The present invention also will be described in further detail in conjunction with the embodiments:
Embodiment one:
With reference to Fig. 1, a kind of occult blood gold standard test strip comprises liner plate 1 and protective seam 2; Be respectively equipped with handheld terminal water accepting layer 3 and the test lead water accepting layer of making by multilayer filter paper 8 at the two ends of liner plate 1; On the liner plate 1 between handheld terminal water accepting layer 3 and the test lead water accepting layer 8, be provided with detection layers 4; Between test lead water accepting layer 8 and detection layers 4, be provided with antihuman hemoglobin gold labeling antibody layer 7; On detection layers 4, be provided with detection line 6 and control line 5.
A kind of preparation method of occult blood gold standard test strip comprises antihuman hemoglobin gold labeling antibody layer 7 and detection layers 4 and goes up the preparation of wrapping tested survey line 6 and control line 5, and the assembling of test-strips.
The preparation of described antihuman hemoglobin gold labeling antibody layer 7 comprises: the preparation of i collaurum, and in being the gold chloride of 0.01-0.03%, concentration adds the trisodium citrate of its volume 1.6%, boiled 10-20 minute, be reduced into the collaurum atom liquid of 15-50 nanometer; Ii antihuman hemoglobin antibody colloidal gold mark adds 6mg antihuman hemoglobin antibody in the 100ml colloidal gold solution; In above-mentioned colloidal gold solution, press 0.24g/100ml again and add bovine serum albumin(BSA); In above-mentioned colloidal gold solution, press the NaCl solution of 10ml/100ml adding 10% then again; After 2000 rev/mins went precipitation in centrifugal 10 minutes, get supernatant again through 10000 rev/mins centrifugal 1 hour sediment, with sediment by 8ml/100ml be dissolved in 0.02M, PH7.4Tris-HCl liquid gets the antihuman hemoglobin colloidal gold solution; Wherein contain 0.25% bovine serum albumin(BSA), 0.02% Sodium azide; Iii is impregnated with glass fibre or nonwoven fabrics with antihuman hemoglobin antibody colloidal gold solution, till the immersion amount begins outwards to flow out with immersion liquid, then at 37 ℃ of dry down golden labeling antibody layers 7 of antihuman hemoglobin that get.
The preparation of described antihuman hemoglobin antibody is that the spleen cell and the rat bone marrow tumour cell that adopt routine techniques will stem from the mouse of using purifying antihuman hemoglobin immunity inoculation merge, and selects to produce the hybridoma of required antihuman hemoglobin monoclonal antibody; Again hybridoma cell clone is got monoclonal antibody.
The preparation of described detection layers 4, in the concentration of preparation is that the antihuman hemoglobin monoclonal antibody of 2mg/ml and concentration are to add 2% fixing agent in the 1mg/ml sheep anti-mouse igg antibody, and Membrane jetter is sprayed on antihuman hemoglobin monoclonal antibody and sheep anti-mouse igg antibody and forms detection line 6 and control line 5 on the cellulose membrane then; Contain 10% calf serum sealing rinsing in 30 minutes, the dry detection layers 4 that gets with 0.01MPH7.0PBS liquid again.Described fixing agent is selected from a kind of in formaldehyde, the acetone.
The last filter paper of pasting handheld terminal water accepting layer 3 and test lead water accepting layer 8 at the two ends of plastics lining board 1 respectively; and section is pasted the detection layers 4 that cellulose membrane is made therein; folder pastes the antibody layer 7 of the glass fibre that contains gold mark antihuman hemoglobin antibody between test lead water accepting layer 8 and detection layers 4; its 4/5 part is clipped in test lead water accepting layer 8; 1/5 part is pressed on the detection layers 4, encloses the protective seam 2 of adhesive sticker then on handheld terminal water accepting layer 3 and test lead water accepting layer 8.
Embodiment two:
Removing colloid gold label antibody is goat-anti human hemoglobin antibody, detection line 6 spray goat-anti human hemoglobin antibody on the cellulose membrane of detection layers 4, outside the anti-sheep IgG of the control line 5 spray rabbits antibody, all the other conditions all with example together.
Embodiment three: removing colloid gold label antibody is rabbit antihuman hemoglobin antibody, detection line 6 spray rabbit antihuman hemoglobin antibody on the cellulose membrane of detection layers 4, outside the control line 5 spray goat anti-rabbit igg antibodies, all the other conditions all with example together.
Claims (6)
1, a kind of occult blood gold standard test strip comprises liner plate (1) and protective seam (2); It is characterized in that being respectively equipped with handheld terminal water accepting layer (3) and test lead water accepting layer (8) at the two ends of liner plate (1); On the liner plate (1) between handheld terminal water accepting layer (3) and the test lead water accepting layer (8), be provided with detection layers (4); Between test lead water accepting layer (8) and detection layers (4), be provided with antihuman hemoglobin antibody gold label antibody layer (7); On detection layers (4), be provided with detection line (6) and control line (5).
2, the preparation method of the described occult blood gold standard test strip of a kind of claim 1, it is characterized in that comprising antihuman hemoglobin antibody gold label antibody layer (7) and detection layers (4) and go up the preparation of wrapping tested survey line (6) and control line (5), and the assembling of test-strips.
3, the preparation method of occult blood gold standard test strip as claimed in claim 2 is characterized in that the preparation of described antihuman hemoglobin antibody gold label antibody layer (7) comprising: the preparation of i collaurum, and adopt known method to prepare collaurum; Ii antihuman hemoglobin antibody colloidal gold mark adds 2-10mg antihuman hemoglobin antibody and comprises monoclonal antibody, sheep rabbit antihuman hemoglobin resist more in the 100ml colloidal gold solution; In above-mentioned colloidal gold solution, press 0.1-0.6g/100ml again and add animal serum albumin; Adding concentration by 6-12ml/100ml again in above-mentioned colloidal gold solution then is 10% saline solution; The last centrifugal precipitation of going, get supernatant centrifugal again sediment, sediment is dissolved in by 4-l0ml/100ml 0.02M, PH7.4Tris-HCl (PBS) contain the 0.1-0.6% animal serum albumin, 0.02% Sodium azide gets antihuman hemoglobin collaurum liquid; Iii is impregnated with glass fibre or nonwoven fabrics with the antihuman hemoglobin colloidal gold solution, till the immersion amount begins outwards to flow out with immersion liquid, then at 37 ℃ of dry down golden labeling antibody layers (7) that get.
4, as the preparation method of occult blood gold standard test strip as described in claim 2 or 3, the preparation that it is characterized in that described antihuman hemoglobin antibody is that the spleen cell and the rat bone marrow tumour cell that adopt routine techniques will stem from the mouse of using purifying antihuman hemoglobin immunity inoculation merge, and selects to produce the hybridoma of required antihuman hemoglobin monoclonal antibody; Again hybridoma cell clone is got monoclonal antibody.
5, the preparation method of occult blood gold standard test strip as claimed in claim 2, it is characterized in that the preparation of described detection layers (4), how anti-add fixing agent at antihuman hemoglobin monoclonal antibody, sheep, rabbit antihuman hemoglobin in and sheep anti mouse, rabbit igg antibody, the anti-sheep IgG of the rabbit antibody, how anti-and sheep anti mouse, rabbit igg antibody, the anti-sheep IgG of rabbit antibody be sprayed on and form detection line (6) and control line (5) on the cellulose membrane with antihuman hemoglobin monoclonal antibody, sheep, rabbit antihuman hemoglobin with Membrane jetter then; Contain 10% calf serum sealing rinsing in 30 minutes, the dry detection layers (4) that gets with 0.01MPH7.0PBS liquid again.
6,, it is characterized in that described fixing agent is selected from a kind of in formaldehyde, the acetone as the preparation method of claim 2 or 5 described occult blood gold standard test strips.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021146497A CN1465977A (en) | 2002-06-21 | 2002-06-21 | Occult blood gold standard test strip and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA021146497A CN1465977A (en) | 2002-06-21 | 2002-06-21 | Occult blood gold standard test strip and preparation method thereof |
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| CN1465977A true CN1465977A (en) | 2004-01-07 |
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| CNA021146497A Pending CN1465977A (en) | 2002-06-21 | 2002-06-21 | Occult blood gold standard test strip and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006092103A1 (en) * | 2005-03-03 | 2006-09-08 | Oakville Hong Kong Co., Limited | Devices and methods for analyte assays with built-in result reporting using recognizable symbols |
| US7704753B2 (en) | 2005-03-03 | 2010-04-27 | Inverness Medical Switzerland Gmbh | Devices and methods for analyte assays with built-in result reporting using recognizable symbols |
| CN101965515A (en) * | 2007-10-30 | 2011-02-02 | 约翰·万 | Methods and device for the detection of occult blood |
| CN1955736B (en) * | 2005-10-28 | 2011-09-14 | 陕西北美基因股份有限公司 | Discrimination antiserum/antibody preparation and its preparation method and use |
| CN101226195B (en) * | 2008-02-03 | 2012-07-11 | 复旦大学附属肿瘤医院 | Kit for detecting knubble type M2 pyruvate kinase and preparation method thereof |
| CN110465335A (en) * | 2018-05-09 | 2019-11-19 | 安徽省立医院 | A kind of micro-fluidic chip and fixture |
-
2002
- 2002-06-21 CN CNA021146497A patent/CN1465977A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006092103A1 (en) * | 2005-03-03 | 2006-09-08 | Oakville Hong Kong Co., Limited | Devices and methods for analyte assays with built-in result reporting using recognizable symbols |
| US7704753B2 (en) | 2005-03-03 | 2010-04-27 | Inverness Medical Switzerland Gmbh | Devices and methods for analyte assays with built-in result reporting using recognizable symbols |
| CN1955736B (en) * | 2005-10-28 | 2011-09-14 | 陕西北美基因股份有限公司 | Discrimination antiserum/antibody preparation and its preparation method and use |
| CN101965515A (en) * | 2007-10-30 | 2011-02-02 | 约翰·万 | Methods and device for the detection of occult blood |
| US8349573B2 (en) | 2007-10-30 | 2013-01-08 | John Wan | Methods and device for the detection of occult blood |
| CN101226195B (en) * | 2008-02-03 | 2012-07-11 | 复旦大学附属肿瘤医院 | Kit for detecting knubble type M2 pyruvate kinase and preparation method thereof |
| CN110465335A (en) * | 2018-05-09 | 2019-11-19 | 安徽省立医院 | A kind of micro-fluidic chip and fixture |
| CN110465335B (en) * | 2018-05-09 | 2021-06-22 | 安徽省立医院 | Micro-fluidic chip and anchor clamps |
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